The homodimerization domain of the Stl repressor is crucial for efficient inhibition of mycobacterial dUTPase
Abstract The dUTPase is a key DNA repair enzyme in Mycobacterium tuberculosis, and it may serve as a novel promising anti-tuberculosis target. Stl repressor from Staphylococcus aureus was shown to bind to and inhibit dUTPases from various sources, and its expression in mycobacterial cells interfered...
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| Format: | Article |
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Nature Portfolio
2024-11-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-024-76349-2 |
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| author | Zoé S. Tóth Ibolya Leveles Kinga Nyíri Gergely N. Nagy Veronika Harmat Thapakorn Jaroentomeechai Oliver Ozohanics Rebecca L. Miller Marina Ballesteros Álvarez Beáta G. Vértessy András Benedek |
| author_facet | Zoé S. Tóth Ibolya Leveles Kinga Nyíri Gergely N. Nagy Veronika Harmat Thapakorn Jaroentomeechai Oliver Ozohanics Rebecca L. Miller Marina Ballesteros Álvarez Beáta G. Vértessy András Benedek |
| author_sort | Zoé S. Tóth |
| collection | DOAJ |
| description | Abstract The dUTPase is a key DNA repair enzyme in Mycobacterium tuberculosis, and it may serve as a novel promising anti-tuberculosis target. Stl repressor from Staphylococcus aureus was shown to bind to and inhibit dUTPases from various sources, and its expression in mycobacterial cells interfered with cell growth. To fine-tune and optimize Stl-induced inhibition of mycobacterial dUTPase, we aimed to decipher the molecular details of this interaction. Structural background of the complex between dUTPase and a truncated Stl lacking the repressor C-terminal homodimerization domain has been described, however, the effects of this truncation of Stl on enzyme binding and inhibition are still not known. Using several independent biophysical, structural and enzyme kinetic methods, here we show that lack of the repressor homodimerization domain strongly perturbs both enzyme binding and inhibition. We also investigated the role of a mycobacteria-specific loop in the Stl-interaction. Our results show that removal of this loop leads to a ten-fold increase in the apparent inhibition constant of Stl. We present a high-resolution three-dimensional structure of mycobacterial dUTPase lacking the genus-specific loop for structural insight. Our present data suggest that potent inhibition of mycobacterial dUTPase by Stl requires the wild-type full-length protein context. |
| format | Article |
| id | doaj-art-60538a93259949f983f23e69b5fb0f06 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-60538a93259949f983f23e69b5fb0f062024-11-10T12:26:54ZengNature PortfolioScientific Reports2045-23222024-11-0114111610.1038/s41598-024-76349-2The homodimerization domain of the Stl repressor is crucial for efficient inhibition of mycobacterial dUTPaseZoé S. Tóth0Ibolya Leveles1Kinga Nyíri2Gergely N. Nagy3Veronika Harmat4Thapakorn Jaroentomeechai5Oliver Ozohanics6Rebecca L. Miller7Marina Ballesteros Álvarez8Beáta G. Vértessy9András Benedek10Institute of Molecular Life Sciences, HUN-REN Research Centre for Natural SciencesInstitute of Molecular Life Sciences, HUN-REN Research Centre for Natural SciencesInstitute of Molecular Life Sciences, HUN-REN Research Centre for Natural SciencesInstitute of Molecular Life Sciences, HUN-REN Research Centre for Natural SciencesLaboratory of Structural Chemistry and Biology, Institute of Chemistry, ELTE Eötvös Loránd UniversityCopenhagen Center for Glycomics, Department of Cellular and Molecular Medicine, University of CopenhagenDepartment of Medical Biochemistry, Semmelweis UniversityCopenhagen Center for Glycomics, Department of Cellular and Molecular Medicine, University of CopenhagenDepartment of Applied Biotechnology and Food Science, Faculty of Chemical Technology and Biotechnology, Budapest University of Technology and EconomicsInstitute of Molecular Life Sciences, HUN-REN Research Centre for Natural SciencesInstitute of Molecular Life Sciences, HUN-REN Research Centre for Natural SciencesAbstract The dUTPase is a key DNA repair enzyme in Mycobacterium tuberculosis, and it may serve as a novel promising anti-tuberculosis target. Stl repressor from Staphylococcus aureus was shown to bind to and inhibit dUTPases from various sources, and its expression in mycobacterial cells interfered with cell growth. To fine-tune and optimize Stl-induced inhibition of mycobacterial dUTPase, we aimed to decipher the molecular details of this interaction. Structural background of the complex between dUTPase and a truncated Stl lacking the repressor C-terminal homodimerization domain has been described, however, the effects of this truncation of Stl on enzyme binding and inhibition are still not known. Using several independent biophysical, structural and enzyme kinetic methods, here we show that lack of the repressor homodimerization domain strongly perturbs both enzyme binding and inhibition. We also investigated the role of a mycobacteria-specific loop in the Stl-interaction. Our results show that removal of this loop leads to a ten-fold increase in the apparent inhibition constant of Stl. We present a high-resolution three-dimensional structure of mycobacterial dUTPase lacking the genus-specific loop for structural insight. Our present data suggest that potent inhibition of mycobacterial dUTPase by Stl requires the wild-type full-length protein context.https://doi.org/10.1038/s41598-024-76349-2 |
| spellingShingle | Zoé S. Tóth Ibolya Leveles Kinga Nyíri Gergely N. Nagy Veronika Harmat Thapakorn Jaroentomeechai Oliver Ozohanics Rebecca L. Miller Marina Ballesteros Álvarez Beáta G. Vértessy András Benedek The homodimerization domain of the Stl repressor is crucial for efficient inhibition of mycobacterial dUTPase Scientific Reports |
| title | The homodimerization domain of the Stl repressor is crucial for efficient inhibition of mycobacterial dUTPase |
| title_full | The homodimerization domain of the Stl repressor is crucial for efficient inhibition of mycobacterial dUTPase |
| title_fullStr | The homodimerization domain of the Stl repressor is crucial for efficient inhibition of mycobacterial dUTPase |
| title_full_unstemmed | The homodimerization domain of the Stl repressor is crucial for efficient inhibition of mycobacterial dUTPase |
| title_short | The homodimerization domain of the Stl repressor is crucial for efficient inhibition of mycobacterial dUTPase |
| title_sort | homodimerization domain of the stl repressor is crucial for efficient inhibition of mycobacterial dutpase |
| url | https://doi.org/10.1038/s41598-024-76349-2 |
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